Tau interactome analyses in CRISPR-Cas9 engineered neuronal cells reveal ATPase-dependent binding of wild-type but not P301L Tau to non-muscle myosins

Abstract

Protein interactions of Tau are of interest in efforts to decipher pathogenesis in Alzheimer’s disease, a subset of frontotemporal dementias, and other tauopathies. We CRISPR-Cas9 edited two human cell lines to generate broadly adaptable models for neurodegeneration research. We applied the system to inducibly express balanced levels of 3-repeat and 4-repeat wild-type or P301L mutant Tau. Following 12-h induction, quantitative mass spectrometry revealed the Parkinson’s disease-causing protein DJ-1 and non-muscle myosins as Tau interactors whose binding to Tau was profoundly influenced by the presence or absence of the P301L mutation. The presence of wild-type Tau stabilized non-muscle myosins at higher steady-state levels. Strikingly, in human differentiated co-cultures of neuronal and glial cells, the preferential interaction of non-muscle myosins to wild-type Tau depended on myosin ATPase activity. Consistently, transgenic P301L Tau mice exhibited reduced phosphorylation of regulatory myosin light chains known to activate this ATPase. The direct link of Tau to non-muscle myosins corroborates independently proposed roles of Tau in maintaining dendritic spines and mitochondrial fission biology, two subcellular niches affected early in tauopathies.

A Complex Containing HNRNPA2B1 and N6-methyladenosine Modified Transcripts Mediates Actions of Toxic Tau Oligomers

“…Comparing tau PPI networks among multiple studies (including our own) identifies tau binding proteins that show up most consistently (Evans et al, 2019;Maziuk et al, 2018;Wang et al, 2019 suggests that oTau-c and oTau share similar patterns of biochemical interactions.…”

Non-Muscle Myosin II in Axonal Cell Biology: From the Growth Cone to the Axon Initial Segment

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Rapid Generation of Human Neuronal Cell Models Enabling Inducible Expression of Proteins-of-interest for Functional Studies

“…Information on statistical analyses as well as were to find publicly available mass spectrometry data from the TAU and BAG5 analyses was included with the article describing the first use of this system (Wang et al, 2019), with Figure 2 of the original article providing details on the design of a quantitative interactome analysis, Figure 3 describing mass spectrometry sequence coverage of the TAU protein, Figure 6 comparing the TAU interactome in IMR32 and ReN VM cells, and Figure 9 documenting results from a post-translational modification analysis of TAU produced with this system. a.…”